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Gram\u2010negative bacteria are the most common organisms causing severe sepsis and responsible for high morbidity and mortality. We aimed to develop a method for rapid multiplex identification of clinically important Gram\u2010negative pathogens and also validated whether our system can identify Gram\u2010negative pathogens with the cell\u2010free plasm DNA from infected blood. We designed five MLPA probe sets targeting the genes specific to major Gram\u2010negative pathogens (<jats:italic>uidA<\/jats:italic> and <jats:italic>lacY<\/jats:italic> for <jats:italic>E. coli<\/jats:italic>, <jats:italic>ompA<\/jats:italic> for <jats:italic>A. baumannii<\/jats:italic>, <jats:italic>phoE<\/jats:italic> for <jats:italic>K. pneumoniae<\/jats:italic>, and <jats:italic>ecfX<\/jats:italic> for <jats:italic>P. aeruginosa<\/jats:italic>) and one set targeting the CTX\u2010M group 1 to identify the ESBL producing Gram\u2010negative pathogens. All six target\u2010specific peaks were clearly separated without any non\u2010specific peaks in a multiplex reaction condition. The minimum detection limit was 100 fg of pathogen DNA. When we tested 28 Gram\u2010negative clinical isolates, all of them were successfully identified without any non\u2010specific peaks. To evaluate the clinical applicability, we tested seven blood samples from febrile patients. Three blood culture positive cases showed <jats:italic>E. coli<\/jats:italic> specific peaks, while no peak was detected in the other four culture negative samples. This technology can be useful for detection of major sepsis\u2010causing, drug\u2010resistant Gram\u2010negative pathogens and also the major ESBL producing Gram\u2010negatives from the blood of sepsis patients in a clinical setting. This system can help early initiation of effective antimicrobial treatment against Gram\u2010negative pathogens for sepsis patients, which is very crucial for better treatment outcomes.<\/jats:p>","DOI":"10.1002\/elps.201700405","type":"journal-article","created":{"date-parts":[[2017,11,29]],"date-time":"2017-11-29T11:10:36Z","timestamp":1511953836000},"page":"645-652","source":"Crossref","is-referenced-by-count":3,"title":["Multiplex identification of sepsis\u2010causing Gram\u2010negative pathogens from the plasma of infected blood"],"prefix":"10.1002","volume":"39","author":[{"given":"Boram","family":"Chung","sequence":"first","affiliation":[{"name":"Department of Microbiology, The Catholic University of Korea College of Medicine  Seoul Republic of Korea"},{"name":"Precision Medicine Research Center, Integrated Research Center for Genome Polymorphism, The Catholic University of Korea College of Medicine  Seoul Republic of Korea"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Chulmin","family":"Park","sequence":"additional","affiliation":[{"name":"Vaccine Bio Research Institute, The Catholic University of Korea, College of Medicine Seoul St. Mary's Hospital  Seoul Republic of Korea"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Sung\u2010Yeon","family":"Cho","sequence":"additional","affiliation":[{"name":"Vaccine Bio Research Institute, The Catholic University of Korea, College of Medicine Seoul St. Mary's Hospital  Seoul Republic of Korea"},{"name":"Department of Internal Medicine, The Catholic University of Korea, College of Medicine Seoul St. Mary's Hospital  Seoul Republic of Korea"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Juyoun","family":"Shin","sequence":"additional","affiliation":[{"name":"Department of Microbiology, The Catholic University of Korea College of Medicine  Seoul Republic of Korea"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Sun","family":"Shin","sequence":"additional","affiliation":[{"name":"Department of Microbiology, The Catholic University of Korea College of Medicine  Seoul Republic of Korea"},{"name":"Precision Medicine Research Center, Integrated Research Center for Genome Polymorphism, The Catholic University of Korea College of Medicine  Seoul Republic of Korea"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Seon\u2010Hee","family":"Yim","sequence":"additional","affiliation":[{"name":"Precision Medicine Research Center, Integrated Research Center for Genome Polymorphism, The Catholic University of Korea College of Medicine  Seoul Republic of Korea"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Dong\u2010Gun","family":"Lee","sequence":"additional","affiliation":[{"name":"Vaccine Bio Research Institute, The Catholic University of Korea, College of Medicine Seoul St. Mary's Hospital  Seoul Republic of Korea"},{"name":"Department of Internal Medicine, The Catholic University of Korea, College of Medicine Seoul St. Mary's Hospital  Seoul Republic of Korea"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6943-5948","authenticated-orcid":false,"given":"Yeun\u2010Jung","family":"Chung","sequence":"additional","affiliation":[{"name":"Department of Microbiology, The Catholic University of Korea College of Medicine  Seoul Republic of Korea"},{"name":"Precision Medicine Research Center, Integrated Research Center for Genome Polymorphism, The Catholic University of Korea College of Medicine  Seoul Republic of Korea"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2017,12,14]]},"reference":[{"key":"e_1_2_8_2_1","doi-asserted-by":"publisher","DOI":"10.1097\/01.CCM.0000194725.48928.3A"},{"key":"e_1_2_8_3_1","doi-asserted-by":"publisher","DOI":"10.1186\/cc1022"},{"key":"e_1_2_8_4_1","doi-asserted-by":"publisher","DOI":"10.1007\/s11908-010-0128-x"},{"key":"e_1_2_8_5_1","doi-asserted-by":"crossref","first-page":"1","DOI":"10.3947\/ic.2014.46.1.1","volume":"46","author":"Cho S. 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